Western Blot Stripping Protocol: Stripping Buffer Recipe (100 mL)1. 12.5 mL of pre-made stripping buffer (0.

25M Tris-HCl, pH 6.8 (62.5 mM) + 16% SDS) 2. 87.5 mL of milliQ H2O 3. 700 L of 2-mercaptoethanol (100 mM) Protocol1. Prepare 50 mL of fresh Stripping buffer for each blot by adding above ingredients. 2. Obtain hybridization chambers from the lab with the hybridization oven (Dr. Tittigers Laboratory). 3. Set the hybridization oven at 50C. 4. Place the blots into the chamber with the protein surface facing inward. 5. Place the assembly into the hybridization oven (*MAKE SURE THE OVEN IS BALANCED they have had trouble and needed to repair it in the past, so handle it respectfully). Do not rotate the apparatus by hand; use the switch to orient it to insert the second chamber. 6. Allow the blots to rotate for 30 minutes. 7. Carefully remove the blots from the oven rotator (make sure to wash and return the glassware to the proper place). 8. Wash the blots three times for 10 minutes with TBS-T or PBS-T. 9. It will be necessary to repeat the blocking step before reprobing the membrane(s).